Vanhove Lenie, Van Gansbeke Thomas, Devriendt Bert, Van der Meeren Ruben, Dmitriev Ruslan I, Okkelman Irina A
Tissue Engineering and Biomaterials Group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, Belgium.
Mitochondrial Investigations Laboratory, Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Belgium.
J Tissue Eng. 2025 Jun 13;16:20417314251345000. doi: 10.1177/20417314251345000. eCollection 2025 Jan-Dec.
Xenogeneic tumour origin and batch-to-batch variability of Engelbreth-Holm-Swarm sarcoma tumour cell-derived hydrogels (Matrigel, Cultrex) limit the biomedical application of organoids in tissue engineering. The gelatin-methacryloyl (GelMA) hydrogels represent a defined, tunable, and GMP-friendly alternative, but they are rarely studied as alternative to Matrigel. Here, we studied effects of mechanical properties of GelMA and addition of laminin-111 on encapsulation and growth of small intestinal organoids. GelMA-embedded organoids displayed polarity reversion, resulting in apical-out and apical-basal phenotypes, independent from the matrix stiffness. Addition of laminin-111 softened hydrogels and also resulted in a partial restoration of the basal-out phenotype. Interestingly, despite the incomplete polarity restoration, GelMA-organoids still showed minor growth. GelMA stiffness and concentration influenced the transition from 3D to 2D organoid cultures. Collectively, our study confirms that tuning of GelMA mechanical properties alone cannot recapitulate the basal membrane matrix. However, controlled polarity reversion offers a tool for engineering organoids and enabling apical membrane access.
异基因肿瘤起源以及Engelbreth-Holm-Swarm肉瘤肿瘤细胞衍生水凝胶（基质胶、Cultrex）的批次间差异限制了类器官在组织工程中的生物医学应用。明胶-甲基丙烯酰基（GelMA）水凝胶是一种明确、可调节且符合药品生产质量管理规范（GMP）要求的替代物，但作为基质胶的替代物却很少被研究。在此，我们研究了GelMA的机械性能以及添加层粘连蛋白-111对小肠类器官包封和生长的影响。嵌入GelMA的类器官表现出极性反转，产生顶端向外和顶端-基底表型，这与基质硬度无关。添加层粘连蛋白-111使水凝胶变软，也导致基底向外表型部分恢复。有趣的是，尽管极性恢复不完全，GelMA类器官仍显示出轻微生长。GelMA的硬度和浓度影响类器官从三维（3D）培养向二维（2D）培养的转变。总体而言，我们的研究证实仅调节GelMA的机械性能无法重现基底膜基质。然而，可控的极性反转提供了一种工程化类器官并实现顶端膜接触的工具。
